Aziz Nabil, Said Tamer, Paasch Uwe, Agarwal Ashok
Reproduction Medicine Unit, Liverpool Women's Hospital, Liverpool, UK.
Hum Reprod. 2007 May;22(5):1413-9. doi: 10.1093/humrep/dem016. Epub 2007 Feb 15.
This study aimed to assess the relationship between apoptosis in human ejaculated spermatozoa, sperm morphology and the novel sperm deformity index (SDI).
Semen specimens from 50 healthy donors were prepared by density-gradient centrifugation followed by incubating the prepared sperm with paramagnetic annexin V-conjugated microbeads and subjecting this to magnetic cell sorting (MACS). The procedure delivers two sperm fractions: annexin-negative (non-apoptotic) and annexin-positive (apoptotic). Activated caspase-3 levels and the integrity of the sperm mitochondrial membrane potential (MMP) were assessed as markers of apoptosis in the annexin-negative and -positive aliquots following MACS. Sperm morphology and the SDI scores were assessed using the strict criteria.
Compared with the apoptotic sperm subpopulations, the non-apoptotic sperm subpopulations had an improved sperm morphology profile as demonstrated by significantly higher proportions of sperm with normal morphology and significantly lower SDI scores and percentages of sperm with acrosomal defects, midpiece defects, cytoplasmic droplet and tail defects. There was a significant correlation between sperm morphology attributes studied and the expressed apoptotic markers - caspase-3 activation and MMP integrity.
Non-apoptotic sperm fractions have morphologically superior quality sperm compared with apoptotic fractions as reflected by significantly lower SDI scores. The study results may support abortive apoptosis, where the apoptotic mechanism of sperm is already triggered prior to ejaculation.
本研究旨在评估人类射出精子中的细胞凋亡、精子形态与新的精子畸形指数(SDI)之间的关系。
对50名健康供体的精液标本进行密度梯度离心制备,然后将制备好的精子与顺磁性膜联蛋白V结合微珠孵育,并进行磁性细胞分选(MACS)。该过程产生两个精子部分:膜联蛋白阴性(非凋亡)和膜联蛋白阳性(凋亡)。在MACS之后,评估膜联蛋白阴性和阳性等分试样中活化的半胱天冬酶-3水平以及精子线粒体膜电位(MMP)的完整性,作为细胞凋亡的标志物。使用严格标准评估精子形态和SDI评分。
与凋亡精子亚群相比,非凋亡精子亚群的精子形态特征有所改善,表现为正常形态精子的比例显著更高,SDI评分以及顶体缺陷、中段缺陷、细胞质滴和尾部缺陷精子的百分比显著更低。所研究的精子形态特征与表达的凋亡标志物——半胱天冬酶-3激活和MMP完整性之间存在显著相关性。
与凋亡部分相比,非凋亡精子部分的精子在形态上质量更优,这表现为SDI评分显著更低。研究结果可能支持流产性凋亡,即精子的凋亡机制在射精前就已被触发。